Veridex LLC, Huntingdon Valley, PA, USA.
Int J Oncol. 2011 Mar;38(3):755-60. doi: 10.3892/ijo.2011.896. Epub 2011 Jan 3.
A validated assay for the enumeration of circulating melanoma cells (CMCs) may facilitate the development of more effective therapies for metastatic melanoma patients. In this study CD146+ cells were immunomagnetically enriched from 7.5 ml of blood. Isolated cells were fluorescently stained with DAPI, anti-molecular weight melanoma-associated antigen (HMW-MAA), anti-CD45 and CD34 and Ki67. CMCs were identified as CD146+, HMW-MAA+, CD45-, CD34-, Ki67-/+ cells. Eighty-eight percent of spiked SK-MEL28 cells in 7.5 ml blood were recovered. In all 55 healthy donors ≤1 CMCs were detected in 7.5 ml of blood. A retrospective analysis was conducted comparing CMC counts and overall survival in 79 blood samples from 44 melanoma patients. CMCs ranged from 0 to 8,042 per 7.5 ml. Two or more CMCs were detected in 18 (23%) of the patients and 30-100% (mean 84%) of the CMCs expressed the proliferation marker Ki67. Patients with ≥2 CMCs per 7.5 ml of whole blood, as compared with the group with <2 CMCs, had a shorter overall survival (2.0 months vs. 12.1 months, P=0.001).
一种经过验证的用于计数循环黑素瘤细胞（CMCs）的检测方法可能有助于开发更有效的转移性黑素瘤患者治疗方法。在这项研究中，从 7.5 毫升血液中免疫磁珠分离出 CD146+细胞。分离的细胞用 DAPI、抗高分子量黑素瘤相关抗原（HMW-MAA）、抗 CD45 和 CD34 以及 Ki67 进行荧光染色。CMCs 被鉴定为 CD146+、HMW-MAA+、CD45-、CD34-、Ki67+/-细胞。在 7.5 毫升血液中，88%的 SK-MEL28 细胞被回收。在所有 55 名健康供体中，在 7.5 毫升血液中检测到≤1 个 CMC。对 44 名黑色素瘤患者的 79 份血液样本进行了回顾性分析，比较了 CMC 计数和总生存率。CMCs 的范围从每 7.5 毫升 0 到 8042 个。18 名（23%）患者中检测到 2 个或更多 CMC，并且 30-100%（平均 84%）的 CMC 表达增殖标志物 Ki67。与每 7.5 毫升全血中<2 个 CMC 的患者相比，每 7.5 毫升全血中≥2 个 CMC 的患者总生存率更短（2.0 个月 vs. 12.1 个月，P=0.001）。
